Parachlamydiaceae are potential emerging pathogens that naturally infect free-living amoebae. Intensive-care patients are highly exposed to aerosols and, consequently, exposed to free-living amoebae and to their intracellular hosts. Thus, we tested intensive-care patients for antibodies to Parachlamydia and determined if serum reactivity was associated with pneumonia. Patients who underwent intubation and were hospitalized in our intensive-care unit were eligible. Clinical data and serum were recorded prospectively. Seventy-three sera taken from 37 intensive-care patients and 100 sera from healthy blood donors were tested for reactivity against Parachlamydia by immunofluorescence. We detected an antibody titer greater than or equal to 1:100 in 5 out of 37 intensive-care unit patients (13.5%), including three seroconversions (8.1%). By contrast, no blood donors were reactive against Parachlamydia (P < 0.001). All patients with serological evidence of a recent exposure to Parachlamydia were trauma patients with head injury and aspiration pneumonia. Moreover, both patients with serological evidence of previous exposure to Parachlamydia were admitted for a cerebral hemorrhage. This serological study suggests that Parachlamydiaceae are associated with aspiration pneumonia in trauma patients admitted to intensive-care units.